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The main obstacle to eradicating HIV-1 from patients is post-integration latency (Finzi et al. , 1999). Antiretroviral treatments target only actively replicating virus, while latent infections that have low or no transcriptional activity remain untreated (Sedaghat et al. , 2007). A combination of antiretroviral treatments with latency-purging strategies may accelerate the depletion of latent reservoirs and lead to a cure (Geeraert et al. , 2008). Current strategies to reactivate HIV-1 from latency include use of prostratin, a non-tumor-promoting phorbol ester (Williams et al. , 2004), BET inhibitors (Filippakopoulos et al. , 2010; Delmore et al. , 2011), and histone deacetylase (HDAC) inhibitors, such as suberoylanilidehydroxamic acid ( i.e. , SAHA or Vorinostat) (Kelly et al. , 2003; Archin et al. , 2009; Contreras et al. , 2009; Edelstein et al. , 2009). As the mechanisms of HIV-1 latency are diverse, effective reactivation may require combinatorial strategies (Quivy et al. , 2002). The following protocol describes a flow cytometry-based method to quantify transcriptional activation of the HIV-1 long terminal repeat (LTR) upon drug treatment. This protocol is optimized for studying latently HIV-1-infected Jurkat (J-Lat) cell lines that contain a GFP cassette. J-Lats that contain a different reporter, for example Luciferase, can be treated with drugs as described but have to be analyzed differently.

Flow Cytometric Analysis of Drug-induced HIV-1 Transcriptional Activity in A2 and A72 J-Lat Cell Lines